2025-07-06
ENA first public
ERC000011
ENA-CHECKLIST
SAMEA114008769
External Id
Frontier Science Center for Stem Cell Research, Tongji University, Shanghai 200092, China.
INSDC center name
2023-07-10T14:53:40Z
INSDC last update
public
INSDC status
ERS15992636
SRA accession
E-MTAB-10585_2:C-H3K27ac
Submitter Id
ArrayExpress
broker name
MC38
cell_line
not collected
collection date
house mouse
common name
Protocols: Tumor cells expressing BFP fluorescent protein were isolated from Rag2-/- mice, Tumor Dissociation Kit (Miltenyi,130-096-730) was used to dissociate the tumor tissues into single cells as the manufacturer’s instructions. ACK lysis buffer (Gibco,492-01) was used to remove the red blood cell. Then Dead Cell Removal kit (Miltenyi,130-090-101) was used to remove dead cells by column in MACS Separator,then Then the tumor cells were resuspended with Facs buffer and BFP+ tumor were sorted at 4℃. CUT&Tag experiment was performed according to the reaction system recommended by Vazyme #TD901-2 follow the instructions. To stop the fragmentation reaction, 10 μl 0.5M EDTA,3 μl 10% SDS and 2.5 μl 20 mg/ml Proteinase K were incubated with mixture overnight at 37℃. 150 μl Tris saturated phenol and 150 μl chloroform were added and centrifuge for 5 min at 16,000 g to collect the supernatant. Add 300 μl chloroform and centrifuge to collect the supernatant, then which was added to 750 μl 100% ethanol in tube on ice, and centrifuge for 15 min at 1,6000 g at 4℃to collect the DNA. Last sample was washed using 100% ethanol, and dissolved in double-distilled water. For library construction, set up the PCR reaction as follows: for a total volume of 50 μL, add 24 μL of DNA,5 μL of ddH2O,10 μL of 5 × TAB buffer, 5μL P5 primer X, 5 μL P7 primer X, and 1 μL of TAE. Mix gently and spin briefly. Then PCR amplification program were carried out,
description
colon carcinoma
disease
wild type genotype
genotype
not collected
geographic location (country and/or sea)
injected into Rag2-deficient mice
growth condition
not applicable
isolate
Mus musculus
organism
Mus musculus
organism
neoplasm
organism part
Mus musculus
scientific_name
C-H3K27ac
title